Oral care compositions and methods

ABSTRACT

Described herein are compositions comprising a MMP-13 inhibitor, and methods of using the same.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. patent application Ser. No.13/518,103, filed 12 Jul. 2012, which is a national stage entry under 35U.S.C. §371 of International Patent Application No. PCT/US2010/061491,filed 21 Dec. 2010, which claims priority to U.S. Provisional PatentApplication No. 61/288,359, filed on 21 Dec. 21 2009, the entireties ofwhich are incorporated herein by reference.

BACKGROUND

Matrix metalloproteinases, which are referred to as MMPs, are anaturally occurring family of calcium- and zinc-dependent endopeptidasesthat are found in most mammals. Over-expression and activation of MMPsor an imbalance between MMPs and inhibitors of MMPs have been suggestedas factors in the pathogenesis of diseases characterized by thebreakdown of extracellular matrix or connective tissues.

The major component of periodontium cementum, periodontal ligament andalveolar bone) is organic matrix. Matrix metalloproteinases (MMPs) areinvolved in remodelling the periodontal matrix. Destructive MMPs degradevarious components of the extracellular matrix both in physiological andpathological conditions. The pathologic overproduction of destructiveMMPs leads to an inappropriate and excessive degradation of matrix. Theoverproduction of destructive MMPs facilitates bone resorption by firstdegrading osteoid (the nonmineralized and newly synthesized bone matrix)and then degrading the matrix, resulting in the clinical manifestationsof periodontitis including gingival recession, pocket formation, loss ofattachment, tooth mobility and tooth loss.

MMP-13 is one of the major destructive MMPs that plays a role indegradation of the extracellular matrix. The level of MMP-13 expressioncorrelates to periodontitis clinical indexes. MMP-13 is detected indiseased periodontal tissue and in gingival crevicular fluid; however,MMP-13 is not detected in healthy oral mucosa. Uitto et al. AmericanJournal of Pathophysiology, 152(6), 1489 (1998).

Matrix metalloproteinase-13 was known as an enzyme responsible for boneresorption and cartilage destruction in rheumatoid arthritis andosteoarthritis. Elevated levels of MMP-13 are also known to exist ingingival crevicular fluid of patients with chronic periodontitis. Inaddition, MMP-13 is known to contribute to both bone and connectivetissue destruction in patients with periodontal diseases. Ilgenli, T. etal. Oral Diseases, 12, 573 (2006).

Currently, antimicrobials, nonsteroidal anti-inflammatory agents(NSAIDs), bisphosphonates and tetracyclines used in the treatment ofperiodontal disease. These agents often do not provide adequatesymptomatic relief and are not believed to alter the natural progressionof the disease. Furthermore, powerful side effects are found with mostall of these therapies. Hence, there is a great need for safe andeffective therapy for these disorders.

Although there are many treatments for various aspects of periodontaldisease, there remains a need to develop an improved oral compositioncomprising active ingredients which target destructive MMPs thatfacilitate bone resorption and cause tissue breakdown. In particular,there is a need to develop improved oral compositions which targetMMP-13, which contributes to both bone resorption and connective tissuedestruction.

SUMMARY

In a first aspect, a method for the inhibition of matrixmetalloproteinase MMP-13 comprising administering to a subject aneffective therapeutic amount of a matrix metalloproteinase MMP-13inhibitor, wherein the matrix metalloproteinase MMP-13 inhibitor isselected from the group consisting of cranberry extract NDM, acetyl ketoβ-boswellic acid, resveratrol, 2-isopropyl-5-methylcyclohexyl2-hydroxyphenylcarbamate, 4-acetamidophenyl2-isopropyl-5-methylcyclohexyl carbonate, and combinations thereof.

The method provides a new mechanism for effectively inhibiting matrixmetalloproteinase MMP-13 and thereby treating or preventing conditionscaused by MMP-13 expression. The present inventors have surprisinglyfound that cranberry extract NDM, acetyl keto β-boswellic acid (AKBBA),resveratrol, 2-isopropyl-5-methylcyclohexyl 2-hydroxyphenylcarbamate,4-acetamidophenyl 2-isopropyl-5-methylcyclohexyl carbonate, andcombinations thereof, effectively inhibit matrix metalloproteinaseMMP-13.

In a second aspect, an oral care composition, the composition comprisingan orally acceptable carrier and a compound selected from the groupconsisting of cranberry extract non-dialyzable material, acetyl ketoβ-boswellic acid, resveratrol, 2-isopropyl-5-methylcyclohexyl2-hydroxyphenylcarbamate, 4-acetamidophenyl2-isopropyl-5-methylcyclohexyl carbonate, and combinations thereof,wherein the compound is present in an amount effective to inhibitMMP-13.

In a third aspect, an oral care composition comprising a compoundselected from the group consisting of cranberry extract non-dialyzablematerial, acetyl keto β-boswellic acid, resveratrol,2-isopropyl-5-methylcyclohexyl 2-hydroxyphenylcarbamate,4-acetamidophenyl 2-isopropyl-5-methylcycloboxyl carbonate, andcombinations thereof for the treatment or prevention of a conditioncaused by MMP-13 expression.

Also, a method of treating degradation of the extracellular matrix, lossof attachment, tooth loss, tooth mobility, pocket formation and boneloss.

DETAILED DESCRIPTION

As used throughout, ranges are used as a shorthand for describing eachand every value that is within the range. Any value within the range canbe selected as the terminus of the range. In addition, all referencescited herein are hereby incorporated by reference in their entireties.In the event of a conflict in a definition in the present disclosure andthat of a cited reference, the present disclosure controls.

Unless otherwise specified, all percentages and amounts expressed hereinand elsewhere in the specification should be understood to refer topercentages by weight. The amounts given are based on the active weightof the material.

As used herein “flavorant” refers to any material or mixture ofmaterials that enhances the taste of a composition.

Compositions

In some embodiments, a composition comprising a therapeuticallyeffective amount of a MMP-13 inhibitor and an orally acceptable carrier.In some embodiments, the MMP-13 inhibitor is selected from the groupconsisting of cranberry extract non-dialyzable material, acetyl ketoβ-boswellic acid, resveratrol, 2-isopropyl-5-methylcyclohexyl2-hydroxyphenylcarbamate, 4-acetamidophenyl2-isopropyl-5-methylcyclohexyl carbonate, and a combination of two ormore thereof.

In some embodiments, the MMP-13 inhibitor is selected from the groupconsisting of cranberry extract non-dialyzable material, acetyl ketoβ-boswellic acid, resveratrol, 2-isopropyl-5-methylcyclohexyl2-hydroxyphenylcarbamate, 4-acetamidophenyl2-isopropyl-5-methylcyclohexyl carbonate, and a combination of two ormore thereof.

In some embodiments, the composition further comprises an anti-plaqueagent, a whitening agent, antibacterial agent, cleaning agent, aflavoring, agent, a sweetening agent, an adhesion agent, a surfactant, afoam modulator, an abrasive, a pH modifying agent, a humectant, a mouthfeel agent, a colorant, a tartar control (anticalculus) agent, afluoride ion source, a saliva stimulating agent, a nutrient, or acombination of two or more thereof.

Some embodiments further comprise a sweetening agent, alcohol, glycerin,sorbitol propylene glycol, polyethylene glycol, a polymer, alkylpolyglycoside (APG), polysorbate, castor oil, or menthol.

In some embodiments, the sweetening agent is saccharin or sodiumsaccharin. In some embodiments, the alcohol is ethanol. In someembodiments, the polymer is POLOXAMER® 407 or PLURONIC® F108 (bothavailable from BASF Corporation). In some embodiments, the polyethyleneglycol is PEG40.

Any orally acceptable natural or synthetic flavorant can be used, suchas flavoring oils, flavoring aldehydes, esters, alcohols, similarmaterials, and combinations thereof. Flavorants include vanillin, sage,marjoram, parsley oil, spearmint oil, cinnamon oil, oil of wintergreen(methylsalicylate), peppermint oil, clove oil, bay oil, anise oil,eucalyptus oil, citrus oils, fruit oils and essences including thosederived from lemon, orange, lime, grapefruit, apricot, banana, grape,apple, strawberry, cherry, pineapple, etc., bean- and nut-derivedflavors such as coffee, cocoa, cola, peanut, almond, etc., adsorbed andencapsulated flavorants, and mixtures thereof. Also encompassed withinflavorants herein are ingredients that provide fragrance and/or othersensory effect in the mouth, including cooling or warming effects. Suchingredients include menthol, menthyl acetate, menthyl lactate, camphor,eucalyptus oil, eucalyptol, anethote, eugenol, cassia, oxanone,[alpha]-irisone, propenyl guaiethol, thymol, linalool, benzaldehyde,cinnamaldehyde, N-ethyl-p-menthan-3-carboxamine,N,2,3-trimethyl-2-isopropylbutanamide, 3-1-menthoxypropane-1,2-diol,cinnamaldehyde glycerol acetal (CGA), methone glycerol acetal (MGA), andmixtures thereof. One or more flavorants are optionally present in atotal amount of about 0.01% to about 5%, optionally in variousembodiments from about 0.05 to about 2%, from about 0.1% to about 2.5%,and from about 0.1 to about 0.5%.

Sweetening agents among those useful herein include dextrose,polydextrose, sucrose, maltose, dextrin, dried invert sugar, mannose,xylose, ribose, fructose, levulose, galactose, corn syrup, partiallyhydrolyzed starch, hydrogenated starch hydrolysate, sorbitol, mannitol,xylitol, maltitol, isomalt, aspartame, neotame, saccharin and saltsthereof, sucralose, dipeptide-based intense sweeteners, cyclamates,dihydrochalcones, and mixtures thereof.

Mouth-feel agents include materials imparting a desirable texture orother feeling during use of the composition. These may includeagglomerated silica particles that are designed to break down withagitation, such as SORBOSIL® BFG series, (e.g., BFG 10, BFG 50, BFG 100,etc.), CBT60S, CBT70, or AC33/43 silicas, commercially available from PQCorporation, Valley Forge, Pa.

Colorants among those useful herein include pigments, dyes, lakes andagents imparting a particular luster or reflectivity such as pearlingagents. In various embodiments, colorants are operable to provide awhite or light-colored coating on a dental surface, to act as anindicator of locations on a dental surface that have been effectivelycontacted by the composition, and/or to modify appearance, in particularcolor and/or opacity, of the composition to enhance attractiveness tothe consumer. Any orally acceptable colorant can be used, including FD&Cdyes and pigments, talc, mica, magnesium carbonate, calcium carbonate,magnesium silicate, magnesium aluminum silicate, silica, titaniumdioxide, zinc oxide, red, yellow, brown and black iron oxides, ferricammonium ferrocyanide, manganese violet, ultramarine, titaniated mica,bismuth oxychloride, and mixtures thereof. One or more colorants areoptionally present in a total amount of about 0.001% to about 20%, forexample about 0.01% to about 10% or about 0.1% to about 5%.

In certain embodiments, the compositions can further comprise anoptional abrasive useful for example as a polishing agent. Any orallyacceptable abrasive can be used, but type, fineness, (particle size) andamount of abrasive should be selected so that tooth enamel is notexcessively abraded in normal use of the composition. Suitable optionalabrasives include silica, for example in the form of precipitated silicaor as admixed with alumina, insoluble phosphates, calcium carbonate, andmixtures thereof. Among insoluble phosphates useful as abrasives areorthophosphates, polymetaphosphates and pyrophosphates. Illustrativeexamples are dicalcium orthophosphate dihydrate, calcium pyrophosphate,calcium pyrophosphate, tricalcium phosphate, calcium polymetaphosphateand insoluble sodium polymetaphosphate.

In certain embodiments, the compositions optionally comprise a tartarcontrol (anticalculus) agent. Tartar control agents among those usefulherein include salts of any of these agents, for example their alkalimetal and ammonium salts: phosphates and polyphosphates (for examplepyrophosphates), polyaminopropanesulfonic acid (AMPS), polyolefinsulfonates, polyolefin phosphates, diphosphonates such asazacycloalkane-2,2-diphosphonates (e.g.,azacycloheptane-2,2-diphosphonic acid), N-methylazacyclopentane-2,3-diphosphonic acid, ethane-1-hydroxy-1,1-diphosphonicacid (EHDP) and ethane-1-amino-1,1-diphosphonate and phosphonoalkanecarboxylic acids. Useful inorganic phosphate and polyphosphate saltsinclude monobasic, dibasic and tribasic sodium phosphates, sodiumtripolyphosphate, tetrapolyphosphate, mono-, di-, tri- and tetrasodiumpyrophosphates, sodium trimetaphosphate, sodium hexametaphosphate andmixtures thereof.

In certain embodiments, the compositions optionally comprise a fluorideion source and useful, for example, as an anti-caries agent. Any orallyacceptable particulated fluoride ion source can be used, includingpotassium, sodium and ammonium fluorides and monofluorophosphates,stannous fluoride, indium fluoride, amine fluorides such as olaflur(N′-octadecyltrimethylendiamine-N,N,N′-tris(2-ethanol)-dihydrofluoride),and mixtures thereof. One or more fluoride ion sources are optionallypresent in an amount providing a clinically efficacious amount ofsoluble fluoride ion to the oral composition.

In certain embodiments, the compositions optionally comprise a salivastimulating agent useful, for example, in amelioration of dry mouth. Anyorally acceptable saliva stimulating agent can be used, includingwithout limitation food acids such as citric, lactic, malic, succinic,ascorbic, adipic, fumaric and tartaric acids, and mixtures thereof. Oneor more saliva stimulating agents are optionally present in salivastimulating effective total amount.

In certain embodiments, the compositions optionally comprise a nutrient.Suitable nutrients include vitamins, minerals, amino acids, and mixturesthereof. Vitamins include Vitamins C and D, thiamine, riboflavin,calcium pantothenate, niacin, folic acid, nicotinamide, pyridoxine,cyanocobalamin, para-aminobenzoic acid, bioflavonoids, and mixturesthereof. Nutritional supplements include amino acids (such asL-tryptophan, methionine, threonine, levocarnitine and L-carnitine),lipotropics such as choline, inositol, betaine, and linoleic acid), andmixtures thereof.

In various embodiments, the oral composition according is notintentionally swallowed, but is rather retained in the oral cavity for atime sufficient to effect the intended utility. In other portableembodiments (such as a lozenge, mint, bead, wafer, liquid formulated fororal application from a small portable nebulizer, liquid formulated fororal application from a small portable drop-generating bottle, or a softpliable tablet), the oral composition is intentionally swallowed,optionally after retention in the oral cavity for a time sufficient toeffect intended utility.

The oral care compositions of the various embodiments preferably are inthe form of a dentifrice. The term “dentifrice” as used throughout thisdescription, denotes a paste, gel, or liquid formulation. The dentifricemay be in any desired form, such as toothpaste; (including deep striped,surface striped, multi-layered, having a gel surround the paste);powder; beads; mouthwash; mouth rinses; lozenge; dental gel; aperiodontal gel; a liquid suitable for painting a dental surface; achewing gum; a dissolvable, partially dissolvable or non-dissolvablefilm or strip; a wafer; a wipe or towelette; an implant; a foam; atroche; a dental floss, liquid formulated for oral application in asmall portable nebulizer (spray bottle), liquid formulated for oralapplication in a small portable drop-generating, bottle, a soft pliabletablet (“chewie”), or any combinations thereof. As used herein, an“orally acceptable carrier” refers to a material or combination ofmaterials that are safe for use in the compositions, commensurate with areasonable benefit/risk ratio.

As used herein the terms “orally acceptable vehicle” or “orallyacceptable carrier” refer to any vehicle useful in formulating any ofthe dentifrices described above. Suitable orally acceptable vehiclesinclude, for example, one or more of the following: a solvent, analkaline agent, a humectant, a thickener, a surfactant, an abrasive, ananti-calculus agent, a colorant, a flavoring agent, a dye, a potassiumcontaining salt, an anti-bacterial agent, desensitizing agents, stainreducing agents, and mixtures thereof.

Some embodiments also provide a composition selected from the groupconsisting of: a lozenge, a mint, a bead, a wafer, a small portablenebulizer containing said admixture in liquid formulated for oralapplication as a spray, a small portable bottle containing saidadmixture in liquid formulated for oral application as a drop, and asoft pliable tablet.

The cranberry extract non-dialyzable material (NDM) is derived fromcranberry juice concentrate. Cranberry juice contains high molecularweight materials (NDM) that inhibit bacterial adhesion to host cells aswell as the co-aggregation of many oral bacteria. The cranberry extractNDM was prepared according to a method described by Weiss E; Lev-Dor,R.; Kashmamn, Y.; Goldhar, J.; Sharon, N.; Ofek, Itzhak, J. Am. Dent.Assoc. 129, 1719 (1998).

Acetyl keto β-boswellic acid (AKBBA) is a useful active compoundisolated from the Boswellia plant which exhibits antibacterial,anti-inflammatory and antioxidant activities. Boswellia is a genus oftrees that produce extracts having anti-inflammatory properties,including boswellic acid compounds. For example, Boswellia sacra, B.frereana; B. serrata; and B. papyrifera and their sub-species producesuitable extracts.

Resveratrol (3,5,4′-trihydroxystilbene), the parent compound of a familyof molecules including glucosides and polymers, is a potent AhRantagonist as described in French Patent Application No. 9805673 filedMay 5, 1998. It is an anti-fungal agent or phytoalexin produced byplants classified as spermatophytes of which vines, peanuts and pinesare prime representatives (Soleas et al., 1997, Clin Biochemistry,30:91-113). As an AhR antagonist, resveratrol, the chemical name ofwhich is 3,5,4′-trihydroxystilbene, is useful generally to prevent thetoxic effects of environmental exposure to AhR ligands, including, forexample, halogenated and polycyclic aryl hydrocarbons, polyaromatichydrocarbons and polychlorinated biphenyls. In addition, resveratrol hasbeen demonstrated to prevent the induction of the proinflammatorycytokine, IL-1 Beta, by AhR ligands (Casper et al. 1999, MolecularPharmacology, 56:784-790).

Methods of Use

Some embodiments provide methods for treating conditions associated withaberrant MMP-13 expression, comprising administering to an animal, inneed thereof, a therapeutically effective amount of a MMP-13 inhibitor.In some embodiments, the composition is suitable for administration orapplication to the oral cavity of an animal.

In some embodiments, provided is the use of a MMP-13 inhibitor in themanufacture of a medicament for treating conditions associated withaberrant MMP-13 expression.

In other embodiments, the MMP-13 inhibitor is selected from the groupconsisting of cranberry extract NDM, acetyl keto β-boswellic acid,resveratrol, 2-isopropyl-5-methylcyclohexyl 2-hydroxyphenylcarbamate and4-acetamidophenyl 2-isopropyl-5-methylcyclohexyl carbonate, or acombination of two or more thereof. In some embodiments, the conditionassociated with MMP-13 expression is selected from periodontal disease,degradation of the extracellular matrix, tooth mobility, tooth decay,loss of attachment, tooth loss, pocket formation or bone loss.

EXAMPLES

Parathyroid hormone (PTH) stimulates the transcription of MMP-13. Theexperimental method utilizes PTH to stimulate MMP-13 transcription inUMR 106-01 cells, which are cells derived from a rat osteoblastic cellline. The UMR 106-01 cell line provides useful model system for studyingeffects of PTH on osteoblasts in vitro. See, e.g., Qin, L. et al.Journal of Biological Chemistry, 278(22), 19723 (2003).

Material

Parathyroid hormone (rat PTH 1-34) was purchased from Sigma.

Cell Culture

The UMR 106-01 cells were cultured in Eagle's minimal essential medium(EMEM) supplemented with 25 mM Hepes 7.4, 1% nonessential amino acids,100 units/ml penicillin, 100 μg/ml streptomycin and 5% fetal bovineserum.

Real Time Quantitative RT-PCR

UMR 106-01 cells were seeded in 12-well plates and cultured for 2-3 daysin cell culture media. When cells were confluent, the cell culture mediawas changed to 1% fetal bovine serum (not 5% fetal bovine serum) forovernight cell starvation. The cells were preincubated with activeingredient for 15 min and then incubated with PTH (10⁻⁸ M) for 4 hours.

Total RNA from UMR 106-01 cells stimulated with or without PTH wasisolated with TRIzol reagent. Total RNA (0.1 μg) was reverse-transcribedto cDNA with the Invitrogen Superscript kit according to themanufacturer's instructions. PCR was performed on cDNA using primers,and the sequences used are listed below. All were amplified by adding2.5 μl of cDNA to the PCR mixture (22.5 μl) containing each primer (0.2μM) and 12.5 μl of the Platinum SYBR Green qPCR SuperMix UDG(Invitrogen). The reactions were pre-incubated at 50° C. for 2 minutesfor decontamination of dU-containing DNA by UDG, then at 95° C. for 2minutes to inactivate UDG and activate Taq. The PCR program continuedfor 49 cycles of denaturation at 95° C. for 15 seconds, annealing andelongation of the primers at 60° C. for 30 seconds. Relativequantification of gene expression was determined by using the 2-deltadelta CT method where fold changes in gene expression are relative tocontrol samples. All samples were normalized to β-actin.

Primer sequence Rat MMP-13 gene 5′-GCCCTATCCCTTGATGCCATT-3′ (sense)5′-ACAGTTCAGGCTCAACCTGCTG-3′ (antisense) Rat β-actin5′-AGCCATGTACGTAGCCATCC-3′ (sense) 5′-ACCCTCATAGATGGGCACAG-3′(antisense)

Example 1

The inhibitory effect of cranberry extract non-dialyzable material(cranberry extract NDM) on PTH stimulated MMP-13 expression was tested.

The cranberry extract NDM was prepared according to a method describedby Weiss, et al. J. Am. Dent. Assoc. 129(12), 1719 (1998). The cranberryextract NDM was obtained by dialyzing cranberry juice through a highmolecular weight cut-off dialysis bag. The substance left in the bagthat does not dialyze out is the non-dialyzable material (NDM).

Cranberry extract NDM at 10 ppm, 4 ppm and 1 ppm in simple solutionshowed MMP-13 inhibition activity. The fold change in MMP-13 geneexpression relative to the control sample is shown in Table 1.

TABLE 1 Average Standard deviation Negative Control 1.07 0.35 PTH 72.882.01 Cranberry 10 ppm 0.78 0.32 Cranberry 4 ppm 1.22 0.15 Cranberry 1ppm 2.57 1.41 PTH + Cranberry 10 ppm 33.03 11.04 PTH + Cranberry 4 ppm46.72 9.27 PTH + Cranberry 1 ppm 54.49 5.86

The results shown in Table 1 demonstrate that cranberry extract NDMdecreases the expression of PTH stimulated MMP-13 gene expression. Thissuggests that cranberry extract NDM prevents matrix degradation byreducing expression of MMP-13.

Example 2

The inhibitory effect of AKBBA on PTH stimulated MMP-13 expression wastested. AKBBA was purchased from Sabinsa Corporation.

AKBBA at 10 ppm in simple solution showed MMP-13 inhibition activity inTable 2. The fold change in MMP-13 gene expression relative to thecontrol sample is shown in Table 2.

TABLE 2 Average Standard deviation Negative Control 0.89 0.25 PTH 406.05122.16 AKBBA 10 ppm 1.45 0.62 PTH + AKBBA 10 ppm 59.77 18.33

The results shown in Table 2 demonstrate that AKBBA decreases theexpression of PTH stimulated MMP-13 gene expression. This suggests thatAKBBA prevents matrix degradation by reducing the expression of MMP-13.

Example 3

Resveratrol (3,5,4′-trihydrostilbene) is a polyphenolic compound foundin grapes, especially in grape skin and seeds. Resveratrol was purchasedfrom Sabinsa Corporation.

Resveratrol at 100 ppm, 50 ppm, 10 ppm, 1 ppm and 0.1 ppm in simplesolution showed MMP-13 inhibition activity (Table 3). The fold change inMMP-13 gene expression relative to the control sample is shown in Table3.

TABLE 3 Average Standard deviation Negative Control 0.68 0.28 PTH 54.646.03 Resveratrol 100 ppm 1.09 0.04 Resveratrol 50 ppm 0.38 0.22Resveratrol 10 ppm 0.65 0.09 Resveratrol 1 ppm 0.71 0.06 Resveratrol 0.1ppm 1.07 0.24 PTH + Resveratrol 100 ppm 0.65 0.24 PTH + Resveratrol 50ppm 0.91 0.79 PTH + Resveratrol 10 ppm 1.86 0.69 PTH + Resveratrol 1 ppm19.97 1.81 PTH + Resveratrol 0.1 ppm 46.02 5.78

The results shown in Table 3 demonstrate that resveratrol decreases theexpression of PTH stimulated MMP-13 gene expression. This suggests thatresveratrol prevents matrix degradation by reducing expression ofMMP-13.

Example 4

Compound A (2-isopropyl-5-methylcyclohexyl 2-hydroxyphenylcarbantate)has a representative structure of Compound A is:

Compound A at 7 ppm in simple solution showed MMP-13 inhibition activity(Table 4). The fold change in MMP-13 gene expression relative to thecontrol sample is shown in Table 4.

TABLE 4 Average Standard deviation Negative control 1.19 0.16 PTH 445.18111.95 Compound A at 7 ppm 7.41 2.65 PTH + Compound A at 7 ppm 14.6743.68

The results shown in Table 4 demonstrate that Compound A decreases theexpression of PTH stimulated MMP-13 gene expression. This suggests thatCompound A prevents matrix degradation by reducing expression of MMP-13.

Example 5

Compound B (4-acetamidophenyl 2-isopropyl-5-methylcyclohexyl carbonate)has a representative structure of Compound B is:

Compound B at 10 ppm in simple solution showed MMP-13 inhibitionactivity Table 5). The fold change in MMP-13 gene expression relative tothe control sample is shown in Table 5.

TABLE 5 Average Standard deviation Negative control 1.24 0.24 PTH 257.0320.82 Compound B at 10 ppm 5.78 1.94 PTH + Compound B at 10 ppm 49.8625.03

The results shown in Table 5 demonstrate that compound B decreases theexpression of PTH stimulated MMP-13 gene expression. Since the level ofMMP-13 is correlated to periodontal clinical indexes, this suggests thatCompound B prevents matrix degradation by reducing MMP-13 withparathyroid hormone (PTH) stimulation.

What is claimed is:
 1. A composition comprising a therapeuticallyeffective amount of a MMP-13 inhibitor, and an orally acceptablecarrier, wherein the MMP-13 inhibitor is 4-acetamidophenyl2-isopropyl-5-methylcyclohexyl carbonate.
 2. The composition of claim 1,wherein the composition further comprises an MMP-13 inhibitor selectedfrom the group consisting of cranberry extract non-dialyzable material,acetyl keto β-boswellic acid, resveratrol, and2-isopropyl-5-methylcyclohexyl 2-hydroxyphenylcarbamate, andcombinations thereof.
 3. The composition of claim 2, wherein the MMP-13inhibitor comprises cranberry extract non-dialyzable material.
 4. Thecomposition of claim 2, wherein the MMP-13 inhibitor comprises acetylketo β-boswellic acid.
 5. The composition of claim 2, wherein the MMP-13inhibitor comprises resveratrol.
 6. The composition of claim 2, whereinthe MMP-13 inhibitor comprises 2-isopropyl-5-methylcyclohex2-hydroxyphenylcarbamate.
 7. The composition of claim 1, wherein thecomposition is a dentifrice.
 8. The composition of claim 7, wherein thedentifrice is selected from the group consisting of: toothpaste; deepstriped toothpaste; surface striped toothpaste; multi-layeredtoothpaste; a gel surrounding toothpaste; powder; beads; mouthwash;mouth rinses; lozenge; dental gel; a periodontal gel; a liquid suitablefor painting a dental surface; a chewing gum; a dissolvable, partiallydissolvable or non-dissolvable film or strip; a wafer; a wipe ortowelette; an implant; a foam; a troche; a dental floss, liquidformulated for oral application in a small portable bottle; liquidformulated for oral application in a small portable drop-generatingbottle; a soft pliable tablet; or a combination of two or more thereof.